Method of making a soft gel capsule comprising coq-10 solubilized in a monoterpene

ABSTRACT

The present invention is directed to compositions and methods of delivery of CoQ-10 solubilized in monoterpenes. Use of monoterpenes as dissolving agents, greatly effects the ability to incorporate greater amounts of bioactive CoQ-10 in formulations, such as soft gel capsules.

CROSS-REFERENCE TO RELATED APPLICATION(S)

This is a Continuation application that claims benefit under 35 U.S.C.§120 to application Ser. No. 10/674,268, filed Sep. 29, 2003, entitled“Solubilized CoQ-10” (Attorney Docket No. 33503/US), the contents ofwhich are incorporated herein in their entirety for all purposes.

FIELD OF THE INVENTION

The present invention relates to the solubilization of coenzyme Q-10 andanalogs thereof in monoterpenes, thereby providing increasedbioavailability in delivery.

BACKGROUND OF THE INVENTION

CoQ-10 (coenzyme Q10) is a fat-soluble quinone that is structurallysimilar to vitamin K and commonly known as ubiquinone. CoQ-10 is foundin most living organisms, and is essential for the production ofcellular energy. CoQ-10 (2,3 dimethyl-5 methyl-6-decaprenylbenzoquinone) is an endogenous antioxidant found in small amounts inmeats and seafood. Although CoQ-10 is found in all human cells, thehighest concentrations of CoQ-10 occur in the heart, liver, kidneys, andpancreas. It is found naturally in the organs of many mammalian species.

CoQ-10 can be synthesized in the body or it can be derived from dietarysources. Situations may arise, however, when the need for CoQ-10surpasses the body's ability to synthesize it. CoQ-10 can be absorbed byoral supplementation as evidenced by significant increases in serumCoQ-10 levels after supplementation.

CoQ-10 is an important nutrient because it lies within the membrane of acell organelle called the mitochondria. Mitochondria are known as the“power house” of the cell because of their ability to produce cellularenergy, or ATP, by shuttling protons derived from nutrient breakdownthrough the process of aerobic (oxygen) metabolism. CoQ-10 also has asecondary role as an antioxidant. CoQ-10, due to the involvement in ATPsynthesis, affects the function of almost all cells in the body, makingit essential for the health of all human tissues and organs. CoQ-10particularly effects the cells that are the most metabolically active:heart, immune system, gingiva, and gastric mucosa

Several clinical trials have shown CoQ-10 to be effective in supportingblood pressure and cholesterol levels. Furthermore, CoQ-10 has also beenshown to improve cardiovascular health. CoQ-10 has been implicated asbeing an essential component in thwarting various diseases such ascertain types of cancers. These facts lead many to believe that CoQ-10supplementation is vital to an individual's well being.

CoQ-10 is sparingly soluble in most hydrophilic solvents such as water.Therefore, CoQ-10 is often administered in a powdered form, as in atablet or as a suspension. However, delivery of CoQ-10 by these methodslimits the bioavailability of the material to the individual.

There is a need in the art for an improved methodology to deliverincreased amount of bioavailable CoQ-10 to an individual in needthereof.

BRIEF SUMMARY OF THE INVENTION

The present invention pertains to the surprising discovery thatubiquinone (CoQ-10) can be readily dissolved in varying concentrationsin monoterpenes. Generally, until the present discovery, most CoQ-10liquid delivery methods could solubilize only up to about 5% by weightof the CoQ-10 in the “solvent”. Typical solvents included various oilsor the CoQ-10 was held in suspension. The present invention provides theability to solubilize CoQ-10 in monoterpenes in concentrations of up toabout 60% (weight to weight) without the need to aggressively heat thesolution or with gentle warming. In particular, the solubilization ofthe CoQ-10 with monoterpenes can be accomplished at ambienttemperatures.

In one aspect, the present invention pertains to compositions thatinclude coenzyme Q-10 or an analog thereof with a sufficient quantity ofa monoterpene that is suitable to solubilize said coenzyme Q-10 and apharmaceutically acceptable carrier. Generally, about 30 to about 45% ofthe CoQ-10 (by weight) is solubilized in the monoterpene. In particular,the monoterpene is limonene. The compositions of the invention areuseful as dietary supplements or as nutriceuticals.

In particular, the compositions of the invention are included in a softgelatin (soft gel) capsule. Typically, the soft gelatin capsule includesat least 5% by weight of coenzyme Q-10 or an analog thereof solubilizedin a monoterpene. Typical monoterpenes include, for example, perillylalcohol, perillic acid, cis-dihydroperillic acid, trans-dihydroperillicacid, methyl esters of perillic acid, methyl esters of dihydroperillicacid, limonene-2-diol, uroterpenol, and combinations thereof.

In another embodiment, the present invention pertains to methods fordelivery of an effective amount of bioavailable CoQ-10 to an individual.The method includes providing CoQ-10 solubilized in a monoterpene, suchthat an effective amount of CoQ-10 is provided to the individual.

In still another embodiment, the present invention also includespackaged formulations of the invention that include a monoterpene as asolvent for the CoQ-10 and instructions for use of the tablet, capsule,elixir, etc.

While multiple embodiments are disclosed, still other embodiments of thepresent invention will become apparent to those skilled in the art fromthe following detailed description, which shows and describesillustrative embodiments of the invention. As will be realized, theinvention is capable of modifications in various obvious aspects, allwithout departing from the spirit and scope of the present invention.Accordingly, the drawings and detailed description are to be regarded asillustrative in nature and not restrictive.

DETAILED DESCRIPTION

The present invention pertains to the surprising discovery thatubiquinone (CoQ-10) can be readily dissolved in varying concentrationsin monoterpenes. CoQ-10 is found in most living organisms, and isessential for the production of cellular energy. Ubiquinone is anaturally occurring hydrogen carrier in the respiratory chain (coenzymeQ) and structurally, it is a 2,3-dimethoxy-5-methyl-1,4-benzoquinonewith a multiprenyl side chain, the number of isoprene units varyingdepending upon the organism from which it is derived. CoQ-10 analogsinclude reduced and semi-reduced CoQ-10 and ubiquinone derivativesdescribed, for example, in WO 8803015, the teachings of which areincorporated herein by reference.

Generally, until the present discovery, most CoQ-10 liquid deliverymethods could solubilize only up at most about 10% by weight of theCoQ-10 in the “solvent. Typical solvents included oils or the CoQ-10 washeld in an aqueous suspension. Alternatively, the CoQ-10 was provided asa solid in a tablet or powder.

The present invention provides the ability to solubilize CoQ-10 inmonoterpenes, as defined herein, in concentrations of up to about 60%(weight to weight) without the need to heat the solution. In one aspect,the monoterpene solubilizes CoQ-10 from about 0.1 percent by weight toabout 45 percent by weight.

In particular, the solubilization of the CoQ-10 with monoterpenes can beaccomplished at ambient temperatures. In one aspect, from about 5 toabout 50 percent (weight CoQ-10/weight solvent) CoQ-10 can besolubilized in a monoterpene. In another aspect, from about 15 to about40 percent w/w can be solubilized and in still another aspect, fromabout 20 to about 35 percent w/w CoQ-10 can be solubilized in amonoterpene.

The phrase “sufficient quantity of a monoterpene suitable to solubilizecoenzyme Q-10” is therefore intended to mean that that amount of amonoterpene that will dissolve CoQ-10 under a given set of conditions,generally, those at ambient temperature. This determination should beunderstood by one skilled in the art and can be determined by methodsknown in the art, such as by solubility studies.

One of the particular advantages of utilizing monoterpenes incombination with CoQ-10 is that the enzyme is dissolved by themonoterpene. That is, many formulations currently in the marketplacehave CoQ-10 present as a suspension; a situation where not all theCoQ-10 is dissolved. This reduces efficacy and the bioavailability ofthe CoQ-10. The present invention eliminates this disadvantage bysolubilizing the CoQ-10 in the monoterpene.

A particular advantage in using monoterpenes is that the CoQ-10 does nothave to be heated to dissolve into solution. This is important so thatthe CoQ-10 does not degrade upon dissolution.

The term “monoterpene” as used herein, refers to a compound having a10-carbon skeleton with non-linear branches. A monoterpene refers to acompound with two isoprene units connected in a head-to-end manner. Theterm “monoterpene” is also intended to include “monoterpenoid”, whichrefers to a monoterpene-like substance and may be used loosely herein torefer collectively to monoterpenoid derivatives as well as monoterpenoidanalogs. Monoterpenoids can therefore include monoterpenes, alcohols,ketones, aldehydes, ethers, acids, hydrocarbons without an oxygenfunctional group, and so forth.

It is common practice to refer to certain phenolic compounds, such aseugenol, thymol and carvacrol, as monoterpenoids because their functionis essentially the same as a monoterpenoid. However, these compounds arenot technically “monoterpenoids” (or “monoterpenes”) because they arenot synthesized by the same isoprene biosynthesis pathway, but rather byproduction of phenols from tyrosine. However, common practice will befollowed herein. Suitable examples of monoterpenes include, but are notlimited to, limonene, pinene, cintronellol, terpinene, nerol, menthane,carveol, S-linalool, safrol, cinnamic acid, apiol, geraniol, thymol,citral, carvone, camphor, etc. and derivatives thereof. For informationabout the structure and synthesis of terpenes, including terpenes of theinvention, see Kirk-Othmer Encyclopedia of Chemical Technology, Mark, etal., eds., 22:709-762 3d Ed (1983), the teachings of which areincorporated herein in their entirety.

In particular, suitable limonene derivatives include perillyl alcohol,perillic acid, cis-dihydroperillic acid, trans-dihydroperillic acid,methyl esters of perillic acid, methyl esters of dihydroperillic acid,limonene-2-diol, uroterpenol, and combinations thereof.

Formulation of the CoQ-10 can be accomplished by many methods known inthe art. For example, the solubilized CoQ-10 can be formulated in asuspension, an emulsion, an elixir, a solution, a caplet that harborsthe liquid, or in a soft gelatin capsule. Often the formulation willinclude an acceptable carrier, such as an oil, or other suspendingagent.

Suitable carriers include but are not limited to, for example, fattyacids, esters and salts thereof, that can be derived from any source,including, without limitation, natural or synthetic oils, fats, waxes orcombinations thereof. Moreover, the fatty acids can be derived, withoutlimitation, from non-hydrogenated oils, partially hydrogenated oils,fully hydrogenated oils or combinations thereof. Non-limiting exemplarysources of fatty acids (their esters and salts) include seed oil, fishor marine oil, canola oil, vegetable oil, safflower oil, sunflower oil,nasturtium seed oil, mustard seed oil, olive oil, sesame oil, soybeanoil, corn oil, peanut oil, cottonseed oil, rice bran oil, babassu nutoil, palm oil, low erucic rapeseed oil, palm kernel oil, lupin oil,coconut oil, flaxseed oil, evening primrose oil, jojoba, tallow, beeftallow, butter, chicken fat, lard, dairy butterfat, shea butter orcombinations thereof.

Specific non-limiting exemplary fish or marine oil sources includeshellfish oil, tuna oil, mackerel oil, salmon oil, menhaden, anchovy,herring, trout, sardines or combinations thereof. In particular, thesource of the fatty acids is fish or marine oil (DHA or EPA), soybeanoil or flaxseed oil. Alternatively or in combination with one of theabove identified carrier, beeswax can be used as a suitable carrier, aswell as suspending agents such as silica (silicon dioxide).

The formulations of the invention are considered dietary supplementsuseful to the increase the amounts of CoQ-10 in the individuals in needthereof.

Alternatively, the formulations of the invention are also considered tobe nutraceuticals. The term “nutraceutical” is recognized in the art andis intended to describe specific chemical compounds found in foods thatmay prevent disease. CoQ-10 is one such compound.

The formulations of the invention can further include variousingredients to help stabilize, or help promote the bioavailability ofthe CoQ-10, or serve as additional nutrients to an individual's diet.Suitable additives can include vitamins and biologically-acceptableminerals. Non-limiting examples of vitamins include vitamin A, Bvitamins, vitamin C, vitamin D. vitamin E, vitamin K and folic acid.Non-limiting examples of minerals include iron, calcium, magnesium,potassium, copper, chromium, zinc, molybdenum, iodine, boron, selenium,manganese, derivatives thereof or combinations thereof. These vitaminsand minerals may be from any source or combination of sources, withoutlimitation. Non-limiting exemplary B vitamins include, withoutlimitation, thiamine, niacinamide, pyridoxine, riboflavin,cyanocobalamin, biotin, pantothenic acid or combinations thereof.

Vitamin(s), if present, are present in the composition of the inventionin an amount ranging from about 5 mg to about 500 mg. More particularly,the vitamin(s) is present in an amount ranging from about 10 mg to about400 mg. Even more specifically, the vitamin(s) is present from about 250mg to about 400 mg. Most specifically, the vitamin(s) is present in anamount ranging from about 10 mg to about 50 mg. For example, B vitaminsare in usually incorporated in the range of about 1 milligram to about10 milligrams, i.e., from about 3 micrograms to about 50 micrograms ofB12. Folic acid, for example, is generally incorporated in a range ofabout 50 to about 400 micrograms, biotin is generally incorporated in arange of about 25 to about 700 micrograms and cyanocobalamin isincorporated in a range of about 3 micrograms to about 50 micrograms.

Mineral(s), if present, are present in the composition of the inventionin an amount ranging from about 25 mg to about 1000 mg. Moreparticularly, the mineral(s) are present in the composition ranging fromabout 25 mg to about 500 mg. Even more particularly, the mineral(s) arepresent in the composition in an amount ranging from about 100 mg toabout 600 mg.

Various additives can be incorporated into the present compositions.Optional additives of the present composition include, withoutlimitation, phospholipids, L-carnitine, starches, sugars, fats,antioxidants, amino acids, proteins, flavorings, coloring agents,hydrolyzed starch(es) and derivatives thereof or combinations thereof.

As used herein, the term “phospholipid” is recognized in the art, andrefers to phosphatidyl glycerol, phosphatidyl inositol, phosphatidylserine, phosphatidyl choline, phosphatidyl ethanolamine, as well asphosphatidic acids, ceramides, cerebrosides, sphingomyelins andcardiolipins.

L-carnitine is recognized in the art and facilitates transport ofmaterials through the mitochondrial membrane. L-carnitine is anessential fatty acid metabolism cofactor that helps to move fatty acidsto the mitochondria from the cytoplasm. This is an important factor asthis is where CoQ-10 uptake occurs.

In one aspect of the present invention, L-carnitine is included in softgel formulations in combination with CoQ-10. Suitable ratios ofL-carnitine and CoQ-10 are known in the art and include those describedin U.S. Pat. No. 4,599,232, issued to Sigma Tau Industrie FaramaceuticheRiunite S.p.A. on Jul. 8, 1986, the teachings of which are incorporatedherein in their entirety. In particular, combinations of limonene,CoQ-10 and L-carnitine in soft gel formulations are of importance. Thepresent invention provides the advantage of solvating large amounts(relative to that of current state of the art) of CoQ-10 in limonene ina soft gel capsule along with an additive, such as L-carnitine.

As used herein, the term “antioxidant” is recognized in the art andrefers to synthetic or natural substances that prevent or delay theoxidative deterioration of a compound. Exemplary antioxidants includetocopherols, flavonoids, catechins, superoxide di smutase, lecithin,gamma oryzanol; vitamins, such as vitamins A, C (ascorbic acid) and Eand beta-carotene; natural components such as camosol, carnosic acid androsmanol found in rosemary and hawthorn extract, proanthocyanidins suchas those found in grapeseed or pine bark extract, and green tea extract.

The term “flavonoid” as used herein is recognized in the art and isintended to include those plant pigments found in many foods that arethought to help protect the body from cancer. These include, forexample, epi-gallo catechin gallate (EGCG), epi-gallo catechin (EGC) andepi-catechin (EC).

Any dosage form, and combinations thereof, are contemplated by thepresent invention. Examples of such dosage forms include, withoutlimitation, chewable tablets, elixirs, liquids, solutions, suspensions,emulsions, capsules, soft gelatin capsules, hard gelatin capsules,caplets, lozenges, chewable lozenges, suppositories, creams, topicals,ingestibles, injectables, infusions, health bars, confections, animalfeeds, cereals, cereal coatings, and combinations thereof. Thepreparation of the above dosage forms are well known to persons ofordinary skill in the art.

For example, health bars can be prepared, without limitation, by mixingthe formulation plus excipients (e.g., binders, fillers, flavors,colors, etc.) to a plastic mass consistency. The mass is then eitherextended or molded to form “candy bar” shapes that are then dried orallowed to solidify to form the final product.

Soft gel or soft gelatin capsules can be prepared, for example, withoutlimitation, by dispersing the formulation in an appropriate vehicle(e.g. rice bran oil, monoterpene and/or beeswax) to form a highviscosity mixture. This mixture is then encapsulated with a gelatinbased film using technology and machinery known to those in the soft gelindustry. The industrial units so formed are then dried to constantweight. Typically, the weight of the capsule is between about 100 toabout 2500 milligrams and in particular weigh between about 1500 andabout 1900 milligrams, and more specifically can weigh between about1500 and about 2000 milligrams.

For example, when preparing soft gelatin shells, the shell can includebetween about 20 to 70 percent gelatin, generally a plasticizer andabout 5 to about 60% by weight sorbitol. The filling of the soft gelatincapsule is liquid (principally limonene, in combination with rice branoil and/or beeswax if desired) and can include, apart form theantioxidant actives, a hydrophilic matrix. The hydrophilic matrix, ifpresent, is a polyethylene glycol having an average molecular weight offrom about 200 to 1000. Further ingredients are optionally thickeningagents. In one embodiment, the hydrophilic matrix includes polyethyleneglycol having an average molecular weight of from about 200 to 1000, 5to 15% glycerol, and 5 to 15% by weight of water. The polyethyleneglycol can also be mixed with propylene glycol and/or propylenecarbonate.

In another embodiment, the soft gel capsule is prepared from gelatin,glycerine, water and various additives. Typically, the percentage (byweight) of the gelatin is between about 30 and about 50 weight percent,in particular between about 35 and about weight percent and morespecifically about 42 weight percent. The formulation includes betweenabout 15 and about 25 weight percent glycerine, more particularlybetween about 17 and about 23 weight percent and more specifically about20 weight percent glycerine.

The remaining portion of the capsule is typically water. The amountvaries from between about 25 weigh percent and about 40 weight percent,more particularly between about 30 and about 35 weight percent, and morespecifically about 35 weight percent. The remainder of the capsule canvary, generally, between about 2 and about 10 weight percent composed ofa flavoring agent(s), sugar, coloring agent(s), etc. or combinationthereof. After the capsule is processed, the water content of the finalcapsule is often between about 5 and about 10 weight percent, moreparticularly 7 and about 12 weight percent, and more specificallybetween about 9 and about 10 weight percent.

As for the manufacturing, it is contemplated that standard soft shellgelatin capsule manufacturing techniques can be used to prepare thesoft-shell product. Examples of useful manufacturing techniques are theplate process, the rotary die process pioneered by R. P. Scherer, theprocess using the Norton capsule machine, and the Accogel machine andprocess developed by Lederle. Each of these processes are maturetechnologies and are all widely available to any one wishing to preparesoft gelatin capsules.

Typically, when a soft gel capsule is prepared, the total weight isbetween about 250 milligrams and about 2.5 gram in weight, e.g., 400-750milligrams. Therefore, the total weight of additives, such as vitaminsand antioxidants, is between about 80 milligrams and about 2000milligrams, alternatively, between about 100 milligrams and about 1500milligrams, and in particular between about 120 milligrams and about1200 milligrams.

For example, a soft gel capsule can be prepared by mixing a 35% solutionof CoQ-10 and limonene (w/w) (e.g., 104 milligrams of CoQ-10 in 193.14milligrams of limonene) with between about 0.01 grams and about 0.4grams (e.g., 0.1 grams) tocopherol, between about 200 grams and about250 grams (e.g., 225 grams) rice bran oil and between about 0.01 gramsand about 0.5 grams betacarotene (e.g. about 0.02 grams). The mixture isthen combined with encapsulated within a gelatin capsule as describedabove.

The present invention also provides packaged formulations of amonoterpene with CoQ-10 and instructions for use of the tablet, capsule,elixir, etc. Typically, the packaged formulation, in whatever form, isadministered to an individual in need thereof that requires and increasein the amount of CoQ-10 in the individual's diet. Typically, the dosagerequirements is between about 1 to about 4 dosages a day.

CoQ-10 has been implicated in various biochemical pathways and issuitable for the treatment of cardiovascular conditions, such as thoseassociated with, for example, statin drugs that effect the body'sability to product coQ-10 naturally. CoQ-10 has also been implicated invarious periodontal diseases. Furthermore, CoQ-10 has been implicated inmitochondrial related diseases and disorders, such as the inability toproduct acetyl coenzyme A, neurological disorders, for example, such asParkinson's disease and, Prater-Willey syndrome.

The following examples are intended to be illustrative only and shouldnot be considered limiting.

EXAMPLES

Formulations of CoQ-10 can be prepared in the following ratios by mixingthe components together and then placing into a soft gel capsule.

Component Example 1 Example 2 CoQ-10 104.09 mg 104.09 mg MixedTocopherols 269.03 mg 269.03 mg (372 IU/g) Rice Bran Oil 176.02 mg —Natural Beta Carotene  10.05 mg  10.05 mg (20% by weight) Yellow Beeswax 20.0 mg — D-limonene — 196.02 mg Total weight   580 mg   580 mg

Example 2 demonstrates that the use of limonene solubilizes CoQ-10without the requirement of beeswax and/or rice bran oil being present.Examples 1 and 2 could be incorporated into soft gel capsules bystandard methods known in the art.

Component Example 3 Example 4 CoQ-10 17.95 g 17.95 g EPAX 2050TG 48.77 g45.49 g D-Limonene 35.70 g 35.70 g 5-67 Tocopherol —  0.86 g (1000 IU/g)

Examples 3 and 4 demonstrate that CoQ-10 can be solubilized in scalablequantities. Additives, such as EPAX 2050 TG (an w-3 oil; 20% EPA/50% DHAas triglycerides, remainder fatty acid/triglycerides; Pronova Biocare)and tocopherols (5-67 Tocopherol; BD Industries) can easily beincorporated into such limonene containing formulations. The resultantmixtures contained approximately 100 mg of CoQ-10 per soft gel capsule.Preparation of the soft gel capsules was accomplished by methods wellknown in the art.

Although the present invention has been described with reference topreferred embodiments, persons skilled in the art will recognize thatchanges may be made in form and detail without departing from the spiritand scope of the invention.

All literature and patent references cited throughout the applicationare incorporated by reference into the application for all purposes.

What is claimed is:
 1. A method to prepare a soft gel capsule,comprising the steps: mixing coenzyme Q-10 with a sufficient quantity ofa monoterpene suitable to solubilize said coenzyme Q-10 and anacceptable carrier to form a composition; and encapsulating saidcomposition in a soft gel capsule.